Venoclara impurity reference substance and preparation method thereof

Abstract

The invention discloses a chemical compound shown in a formula (I) of a Venoclara impurity reference substance and a preparation method of the chemical compound. The preparation method comprises the following steps: reacting a compound shown as a formula (II) with a compound shown as a formula (III) in the presence of a proper solvent, an activating agent and a condensing agent to obtain a compound shown as a formula (I), preparing target impurities by adopting a condensation reaction, reducing interference of other reactants, selecting a proper solvent, controlling the proportion and temperature of the reactants, and giving enough reaction time. The reaction efficiency and the conversion rate can be remarkably improved, and finally, the target product is efficiently produced.

Description

Technical Field

[0001] The invention relates to the technical field of chemical drug analysis and drug synthesis, in particular to a venetoclax impurity reference substance and a preparation method thereof. Background Art

[0002] Venetoclax (Venetoclax), CAS registration number: 1257044-40-8, chemical name: 2-((1H-pyrrolo[2,3-B]pyridin-5-yl)oxy)-4-(4-((4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide, its structural formula is:

[0003]

[0004] Venetoclax is used to treat acute myeloid leukemia and chronic lymphocytic leukemia. It can prevent apoptosis of some cells (including lymphocytes) and is overexpressed in certain types of cancer, which is associated with the formation of drug resistance. Venetoclax is designed to selectively inhibit the function of BCL-2, restore the cell's communication system, and allow cancer cells to self-destruct, thereby achieving the purpose of treating tumors.

[0005] At present, the synthesis route of venetoclax is mostly based on 2-fluoro-4-bromo-1-iodobenzene as the initial raw material, and venetoclax is obtained through Grignard reaction, nucleophilic reaction, Buchwald coupling, hydrolysis, and condensation reaction. The synthesis route is as follows:

[0006]

[0007] However, the venetoclax product contains a variety of impurities. At present, technicians need such impurities to enrich and improve the impurity spectrum of venetoclax, and at the same time, the comprehensive control of the quality of venetoclax is of great significance. Such impurities are usually separated from the product, but the obtained impurities have low purity and the separation process is complicated, causing inconvenience.

[0008] It should be noted that the information disclosed in the above background technology section is only used to enhance the understanding of the background of the present disclosure, and therefore may include information that does not constitute the prior art known to ordinary technicians in the field. Summary of the invention

[0009] In view of the deficiencies in the prior art, the invention discloses a venetoclax impurity reference substance and a preparation method thereof.

[0010] The technical solution adopted by the present invention is as follows:

[0011] A method for preparing a venetoclax impurity reference substance, characterized in that it comprises the following steps:

[0012] Compound (II) and compound (III) are combined with a solvent, an activator and a condensing agent to undergo a condensation reaction to obtain impurity (I), reaction formula:

[0013]

[0014] Furthermore, the molar ratio of the compound (II), the compound (III), the solvent, the activating agent and the condensing agent is 1:1-3:1.1-3:1.2-3.

[0015] Furthermore, the solvent is any one of dichloromethane, dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide or tetrahydrofuran, or a mixture of several of them.

[0016] Furthermore, the condensing agent is one of N,N'-dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl).

[0017] Furthermore, the activator is one of 4-dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt), N-hydroxysuccinimide (HOSU), and pentafluorophenol (PFP-OH).

[0018] Furthermore, the condensation reaction temperature is 10-50°C.

[0019] Furthermore, the reaction time is controlled within 6 to 24 hours.

[0020] Furthermore, after the condensation reaction is completed, the reaction product is washed, and the organic phase is dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude impurity (I), and after the impurity (I) is crystallized, a solid impurity (I) is obtained.

[0021] Furthermore, the reaction product is first washed once with 100 mL of 5% acetic acid aqueous solution, and then washed once with 100 mL of 5.0% sodium bicarbonate solution. The organic phase after washing is dried with 20 g of anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude impurity (I). The crude impurity (I) is crystallized using a dichloromethane / methanol system.

[0022] A structure of a Venetoclax impurity is shown in Formula 1:

[0023]

[0024] The beneficial effects of the present invention are as follows:

[0025] 1. Use condensation reaction to prepare target impurities, reduce interference from other reactants, have fewer related side reactions, directly react to obtain the target product, reduce interference from other products, and greatly improve the purity and yield of the target product.

[0026] 2. By selecting a suitable solvent, controlling the ratio of reactants and temperature, and giving enough reaction time, the efficiency and conversion rate of the reaction can be significantly improved. The reaction is carried out at room temperature and the temperature is controlled between 20 and 30 ° C. This temperature range is conducive to the smooth progress of the reaction and avoids side reactions caused by too high temperature or too slow reaction rate caused by too low temperature.

[0027] 3. After the reaction is completed, acetic acid of appropriate concentration is first used to help neutralize the remaining alkali and remove impurities that can react with the acid. Then, sodium bicarbonate solution of appropriate concentration is used to remove acidic impurities, impurities, unreacted substances, catalyst residues and possible by-products in the reaction mixture to improve product purity. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Schematic diagram of the synthetic route of the compound of formula (I).

[0029] Figure 2 It is the HPLC spectrum of the compound of formula (I).

[0030] Figure 3 is the MS spectrum of the compound of formula (I).

[0031] Figure 4 is the 13C NMR spectrum of the compound of formula (I).

[0032] Figure 5 1HNMR spectrum of the synthetic method of venetoclax intermediate. DETAILED DESCRIPTION

[0033] The specific implementation of the present invention will be described below in conjunction with the accompanying drawings.

[0034] Embodiment 1:

[0035] A method for preparing a venetoclax impurity reference substance comprises the following steps:

[0036] Add compound (II) (20.0 g, 35.0 mmol, 1.0 eq) to a three-necked reaction flask, dissolve it in 300 mL of dichloromethane (DCM), and add compound (III) (11.1 g, 38.0 mmol, 1.0 eq), 4-dimethylaminopyridine (DMAP) (5.13 g, 42.0 mmol, 1.2 eq), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) (7.4 g, 38.5 mmol, 1.1 eq) in sequence at room temperature. Control the temperature at 20-30°C and stir the reaction for 16 hours. After the reaction was completed, the temperature was lowered, 100 mL of 5% acetic acid aqueous solution was added for washing once, and then 100 mL of 5.0% sodium bicarbonate solution was used for washing once. The organic phase was dried over 20 g of anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude impurity (I). The crude impurity (I) was then crystallized using a dichloromethane / methanol system to obtain 6.01 g of a solid impurity (I) with a purity of 99.5% and a yield of 19.7%.

[0037] in:

[0038] Compound (II), name: 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((2-(4-chlorophenyl)-4,4-dimethylcyclohexene-1-enyl)methyl)piperazin-1-yl)benzoic acid.

[0039] Properties: White powdery solid, easily soluble in organic solvents such as dichloromethane, insoluble in water, odorless.

[0040] CAS: 1235865-77-6.

[0041] Compound (III), name: 1-[[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-enyl]methyl]piperazine.

[0042] Molecular formula: C19H27ClN2.

[0043] Molecular weight: 318.88.

[0044] [Density]: 1.1±0.1g / cm 3 .

[0045] [Boiling point]: 421.2±45.0℃at 760mmHg.

[0046] [Molecular formula]: C19H27ClN 2 .

[0047] [Molecular weight]: 318.884.

[0048] [Flash point]: 208.5±28.7℃.

[0049] [Exact mass]: 318.186279.

[0050] [Vapor pressure]: 0.0±1.0mmHg at 25℃.

[0051] [Refractive index]: 1.545.

[0052] Impurity (I), this product is a yellow powdery solid with a molecular formula of C 52 H 60 Cl 2 N 6 O 2 , molecular weight 871.99, easily soluble in dichloromethane, insoluble in water, the specific structure is as follows:

[0053]

[0054] The reaction process is as follows Figure 1 shown.

[0055] Final material testing:

[0056] The HPLC spectrum of the white solid is as follows Figure 2 shown.

[0057] The MS spectrum of the white solid is as follows Figure 3 shown.

[0058] White solid 13 C NMR spectrum Figure 4 shown.

[0059] White solid 1 HNMR spectrum Figure 5 shown.

[0060] In summary, the diffraction peak data of the final product are consistent with the characteristics of the designed product.

[0061] Embodiment 2:

[0062] A method for preparing a venetoclax impurity reference substance comprises the following steps:

[0063] Add compound (II) (20.0 g, 35.0 mmol, 1.0 eq) to a three-necked reaction flask, dissolve it in 300 mL of dichloromethane (DCM), and add compound (III) (22.2 g, 70.0 mmol, 2.0 eq), 4-dimethylaminopyridine (DMAP) (6.41 g, 52.5 mmol, 1.5 eq), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) (10.06 g, 52.5 mmol, 1.5 eq) in sequence at room temperature. Control the temperature at 30-40°C and stir the reaction for 12 hours. After the reaction was completed, the temperature was lowered, 100 mL of 5% acetic acid aqueous solution was added for washing once, and then 100 mL of 5.0% sodium bicarbonate solution was used for washing once. The organic phase was dried over 20 g of anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude impurity (I). The crude impurity (I) was then crystallized using a dichloromethane / methanol system to obtain 4.63 g of a solid impurity (I) with a purity of 97.2% and a yield of 15.2%.

[0064] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0065] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for preparing a venetoclax impurity reference substance, characterized in that: The following steps are involved: Compound (II) and compound (III) are combined with a solvent, an activator and a condensing agent to undergo a condensation reaction to obtain impurity (I), reaction formula:

2. The method for preparing the venetoclax impurity reference substance according to claim 1, wherein: The molar ratio of the compound (II), the compound (III), the solvent, the activating agent and the condensing agent is 1:1-3:1.1-3:1.2-3.

3. The method for preparing the venetoclax impurity reference substance according to claim 2, wherein: The solvent is any one of dichloromethane, dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide or tetrahydrofuran, or a mixture of the two or more thereof.

4. The method for preparing the venetoclax impurity reference substance according to claim 2, wherein: The condensing agent is one of N,N'-dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl).

5. The method for preparing the venetoclax impurity reference substance according to claim 2, wherein: The activator is one of 4-dimethylaminopyridine (DMAP), 1-hydroxybenzotriazole (HOBt), N-hydroxysuccinimide (HOSU), and pentafluorophenol (PFP-OH).

6. The method for preparing a venetoclax impurity reference substance according to claim 1, characterized in that: The condensation reaction temperature is 10-50°C.

7. The method for preparing a venetoclax impurity reference substance according to claim 1, characterized in that: The reaction time is controlled within 6 to 24 hours.

8. The method for preparing the venetoclax impurity reference substance according to claim 1, characterized in that: After the condensation reaction is completed, the reaction product is washed, and the organic phase is dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude impurity (I). After the impurity (I) is crystallized, a solid impurity (I) is obtained.

9. The method for preparing the venetoclax impurity reference substance according to claim 8, characterized in that: The reaction product is first washed once with 100 mL of 5% acetic acid aqueous solution, and then washed once with 100 mL of 5.0% sodium bicarbonate solution. The organic phase after washing is dried with 20 g of anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude impurity (I). The crude impurity (I) is crystallized using a dichloromethane / methanol system.

10. A venetoclax impurity prepared by the method for preparing a venetoclax impurity reference substance according to any one of claims 1 to 9, characterized in that: The structure of impurity (I) is shown in Formula 1:

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